The white-rot fungus Phanerochaete sordida YK-624 is capable of degrading various emerging contaminants (ECs), with cytochrome P450 (CYP) enzymes playing crucial catalytic roles in the degradation process. In this study, we first identified 214 putative CYPs in P. sordida YK-624 (PsCYPs), of which 208 PsCYPs were classified into 31 CYP families, and 6 PsCYPs remained unclassified. To construct a comprehensive functional screening system for identifying PsCYPs involved in EC degradation, a heterologous co-expression system was established using Saccharomyces cerevisiae. The heterologous expressing yeasts were then used to evaluate the degradation of 7-ethoxycoumarin, a natural compound commonly used to assess CYP activity. Several expressing yeasts catalyzed the hydroxylation and O-deethylation of 7-ethoxycoumarin, confirming that these yeasts expressed active forms of PsCYPs. Subsequent degradation experiments were conducted on ECs such as carbazole, acetamiprid (ACE), bisphenol A (BPA), and loxoprofen (LOX). Metabolite analyses using LC/MS, GC/MS and NMR revealed that the PsCYPs catalyzed the hydroxylation of carbazole, BPA, and LOX, as well as the N-dealkylation of ACE. These findings not only provide strong evidence supporting our previous finding regarding degradation research mediated by P. sordida YK-624 mycelia, but also offer valuable insights for future bioprospecting of fungal CYPs in the bioremediation of EC.